Ngna compositions and methods of use

ABSTRACT

The present invention relates to the field of viral disorders, and in particular to the use of natural compounds to inhibit viruses and viral infection. Compositions comprising NGNA are provided for treating or preventing viral infections, such as those causing the common cold.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/720,107, filed Dec. 19, 2012, now U.S. Pat. No. 8,628,793, which is acontinuation of abandoned U.S. patent application Ser. No. 12/615,818,filed Nov. 10, 2009, which claims priority to expired U.S. ProvisionalPatent Application Ser. No. 61/113,048, filed Nov. 10, 2008, thecontents of which are herein incorporated by reference in theirentireties.

FIELD OF THE INVENTION

The present invention relates to the use of compositions comprisingn-glycolylneuraminic acid (NGNA). Compositions comprising NGNA find usein inducing physiological responses such as alleviating the symptoms ofcolds and preventing the onset of colds.

BACKGROUND OF THE INVENTION

In the course of a year, individuals in the United States suffer 1billion colds, according to some estimates. Colds are most prevalentamong children, and seem to be related to youngsters' relative lack ofresistance to infection and to contacts with other children in day-carecenters and schools. Children have about six to ten colds a year. Infamilies with children in school, the number of colds per child can beas high as 12 a year. Adults average about two to four colds a year,although the range varies widely. Women, especially those aged 20 to 30years, have more colds than men, possibly because of their closercontact with children. On average, individuals older than 60 have fewerthan one cold a year.

The economic impact of the common cold is enormous. The National Centerfor Health Statistics (NCHS) estimates that, in 1994, 66 million casesof the common cold in the United States required medical attention orresulted in restricted activity. In 1994, colds caused 24 million daysof restricted activity and 20 million days lost from school, accordingto NCHS.

More than 200 different viruses are known to cause the symptoms of thecommon cold. Some, such as the rhinoviruses, seldom produce seriousillnesses. Others, such as parainfluenza and respiratory syncytialvirus, produce mild infections in adults but can precipitate severelower respiratory infections in young children. Rhinoviruses (from theGreek rhin, meaning “nose”) cause an estimated 30 to 35 percent of alladult colds, and are most active in early fall, spring and summer. Morethan 110 distinct rhinovirus types have been identified. These agentsgrow best at temperatures of 33 degrees Celsius [about 91 degreesFahrenheit (F)], the temperature of the human nasal mucosa.Coronaviruses are believed to cause a large percentage of all adultcolds. They induce colds primarily in the winter and early spring. Ofthe more than 30 isolated strains, three or four infect humans. Theimportance of coronaviruses as causative agents is hard to assessbecause, unlike rhinoviruses, they are difficult to grow in thelaboratory.

In the United States, most colds occur during the fall and winter.Beginning in late August or early September, the incidence of coldsincreases slowly for a few weeks and remains high until March or April,when it declines. The seasonal variation may relate to the opening ofschools and to cold weather, which prompt people to spend more timeindoors and increase the chances that viruses will spread from person toperson.

Symptoms of the common cold usually begin two to three days afterinfection and often include nasal discharge, obstruction of nasalbreathing, swelling of the sinus membranes, sneezing, sore throat,cough, and headache. Fever is usually slight but can climb to 102° F. ininfants and young children. Cold symptoms can last from two to 14 days,but two-thirds of people recover in a week. If symptoms occur often orlast much longer than two weeks, they may be the result of an allergyrather than a cold. Colds occasionally can lead to secondary bacterialinfections of the middle ear or sinuses, requiring treatment withantibiotics. High fever, significantly swollen glands, severe facialpain in the sinuses, and a cough that produces mucus, may indicate acomplication or more serious illness requiring a doctor's attention.

Viruses cause infection by overcoming the body's complex defense system.The body's first line of defense is mucus, produced by the membranes inthe nose and throat. Mucus traps inhaled materials: pollen, dust,bacteria and viruses. When a virus penetrates the mucus and enters acell, it commandeers the protein-making machinery to manufacture newviruses which, in turn, attack surrounding cells.

Cold symptoms are probably the result of the body's immune response tothe viral invasion. Virus-infected cells in the nose send out signalsthat recruit specialized white blood cells to the site of the infection.In turn, these cells emit a range of immune system chemicals such askinins. These chemicals probably lead to the symptoms of the common coldby causing swelling and inflammation of the nasal membranes, leakage ofproteins and fluid from capillaries and lymph vessels, and the increasedproduction of mucus. Kinins and other chemicals released by immunesystem cells in the nasal membranes are the subject of intensiveresearch. Researchers are examining whether drugs to block them, or thereceptors on cells to which they bind, might benefit people with colds.

Despite the economic and health costs associated with colds, there arevery few drugs or natural compounds that have been proven to fight theviruses that cause colds. What is needed in the art are naturalcompounds for preventing and treating colds.

SUMMARY OF THE INVENTION

The present invention relates to the use of compositions comprisingn-glycolylneuraminic acid (NGNA). Compositions comprising NGNA find usein inducing physiological responses such as alleviating the symptoms ofviral-induced obesity, alleviating the symptoms of colds, preventing theonset of colds, increasing energy and increasing the feeling ofwell-being in subjects.

In some embodiments, the present invention provides compositions fortreating or preventing colds comprising an effective concentration ofn-glycolylneuraminic acid. In some embodiments, the compositions furthercomprise an excipient. In some embodiments, the compositions areformulated for oral administration. In some embodiments, thecompositions are formulated as a tablet. In some embodiments, thecompositions are formulated as a capsule. In some embodiments, thecompositions further comprise a foodstuff. In some embodiments, thecompositions are formulated as a nasal sprays for nasal administration.In some embodiments, the compositions are gels. In some embodiments, thecompositions are aqueous solutions. In some embodiments, thecompositions are formulated as a pump spray, aerosol spray, gel, powder,lotion, foam or cream for topical or intranasal administration. In someembodiments, the effective concentration is from about 1 nM to about 10mM.

In some embodiments, the present invention provides methods for treatinga human or animal subject comprising providing a composition comprisingn-glycolylneuraminic acid in an effective concentration andadministering said composition intranasally. In some embodiments, thecomposition is selected from the group consisting of gels, solutions,sprays, powders, creams, foams and lotions. In some embodiments, theeffective concentration is from about 1 nM to about 10 mM.

In some embodiments, the present invention provides compositions fortreating or preventing flu comprising an effective concentration ofn-glycolylneuraminic acid to destroy or inhibit influenza virus. In someembodiments, the compositions further comprise an excipient. In someembodiments, the compositions are formulated for oral administration. Insome embodiments, the compositions are formulated as a tablet. In someembodiments, the compositions are formulated as a capsule. In someembodiments, the compositions further comprise a foodstuff. In someembodiments, the compositions are formulated as a nasal sprays for nasaladministration. In some embodiments, the compositions are gels. In someembodiments, the compositions are aqueous solutions. In someembodiments, the compositions are formulated as a pump spray, aerosolspray, gel, powder, lotion, foam or cream for topical or intranasaladministration. In some embodiments, the effective concentration is fromabout 1 nM to about 10 mM. In some embodiments, the influenza virus isselected from the group consisting of Influenza A, B, and C viruses. Insome embodiments, the Influenza A virus has a serotype selected from thegroup consisting of H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2,H7N3, and H10N7 serotypes.

In some embodiments, the present invention provides methods for treatingor preventing flu in a human or animal subject comprising providing acomposition comprising n-glycolylneuraminic acid in an effectiveconcentration and administering said composition under conditions suchthat influenza virus is destroyed or inhibited. In some embodiments, thecomposition is administered intranasally. In some embodiments, thecomposition is selected from the group consisting of gels, solutions,sprays, powders, creams, foams and lotions. In some embodiments, theeffective concentration is from about 1 nM to about 10 mM. In someembodiments, the influenza virus is selected from the group consistingof Influenza A, B, and C viruses. In some embodiments, the Influenza Avirus has a serotype selected from the group consisting of H1N1, H2N2,H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, and H10N7 serotypes.

In some embodiments, the present invention provides methods ofinhibiting a virus comprising contacting said virus with an effectiveconcentration of n-glycolylneuraminic acid, wherein said virus isselected from the group consisting of influenza viruses, rhinoviruses,adenoviruses, herpes viruses and noroviruses. In some embodiments, theeffective concentration is from about 1 nM to about 10 mM.

In some embodiments, the present invention provides an animal feedcomprising an effective concentration of n-glycolylneuraminic acid.

In some embodiments, the present invention provides a device comprisinga solid support having NGNA deposited thereon, wherein said device isconfigured to cover a body cavity on an animal or human.

In some embodiments, the present invention provides a device foradministering an antiviral solution comprising a container containing asolution comprising NGNA and a propellant.

In some embodiments, the present invention provides a device foradministering an antiviral solution comprising NGNA comprising acontainer containing a solution comprising NGNA, a pump and a nozzle,wherein activation of said pump causes said solution comprising NGNA tobe expelled through said nozzle.

In some embodiments, the present invention provides a method fortreating a site suspected of being contaminated by rhinovirus comprisingexposing said site to a composition comprising NGNA under conditionssuch that the percentage of viable or infectious rhinovirus isdecreased. In some embodiments, the site is a surface. In someembodiments, the site is an enclosed space. In some embodiments, theenclosed space is a room. In some embodiments, the enclosed space is avehicle. In some embodiments, the composition comprising NGNA isprovided as an aerosol spray. In some embodiments, the compositioncomprising NGNA is provided as a foam.

In some embodiments, the present invention provides an NGNA compositionfor use in treating or preventing infection by influenza virus. In someembodiments, the NGNA composition is provided as an eyedrop, gel,solution, pump spray, aerosol spray, powder, cream, foam or lotion. Insome embodiments, the present invention contemplates use of NGNA for themanufacture of a medicament for treating or preventing infection byinfluenza virus.

DESCRIPTION OF THE FIGURES

FIG. 1 a provides a chart showing body composition before (For) andafter (etter) the eight week NGNA sport drink study. FIG. 1 b is a graphshowing a decrease in average weight of the subjects involved in thestudy.

FIGS. 2 a-f provide graph of responses over the eight week (Uke) periodof the NGNA sport drink study to the following questions: FIG. 2 a—Howwas your workout today?; FIG. 2 b—How do you feel?; FIG. 2 c: Do youhave a cold 0, or are you healthy 9?; FIG. 2 d: Do you feel the drinkhelps you? FIG. 2 e—How is workout energy and joy?; and FIG. 2 f—Do youlike the sport drink?. The test subject were asked to rate theirresponses to these questions on a scale of 0-9, 9 being the mostpositive.

FIG. 3 is graph of Clinical Signs vs. Days Post Infection.

FIG. 4 is a graph of % Feed Intake Change vs. Days Post Infection.

FIG. 5 is a graph of % Mouse Weight Change vs Days Post Infection.

FIG. 6 is a graph of Lung Consolidation Scores vs Days Post Infection.

FIG. 7 is a graph of Histopathology vs Days Post Infection.

DEFINITIONS

As used herein, the term “obesity” and related terms refers to anincrease in body weight beyond the limitation of skeletal and physicalrequirement, as the result of an excessive accumulation of fat in thebody.

As used herein, the term “anorectic agent” and related terms refer topharmaceutical agents used to induce weight loss in a subject. Examplesof anorectic agents include, but are not limited to, phentermine,phenylpropanolaminea, mazindol, ephedra, and sibutramine.

As used herein, the term “phytonutrient” refers to organic compoundsisolated from plants that have a biological effect, and includes, but isnot limited to, compounds of the following classes: isoflavonoids,oligomeric proanthcyanidins, indol-3-carbinol, sulforaphone, fibrousligands, plant phytosterols, ferulic acid, anthocyanocides, triterpenes,omega 3/6 fatty acids, polyacetylene, quinones, terpenes, cathechins,gallates, and quercitin.

As used herein, the term “functional foods” refers to food products thatinclude biologically active nutraceutical agents.

As used herein, the terms “nutraceutical agent,” and related terms,refer to natural, bioactive chemical compounds that have healthpromoting, disease preventing or medicinal properties. Examples ofnutraceutical agents include, but are not limited to, Allium Cepa,Allium Sativum, Aloe Vera, Angelica Species, Naturally OccurringAntioxidants, Aspergillus Oryzae Enzyme Therapy, barley grass,Bromelain, Carnitine, Carotenoids and Flavonoids, Catechin, CentellaAsiatica (Gotu kola), Coenzyme Q10, Chinese Prepared Medicines, ColeusForskohlii, Commiphora Mukul, Crataegus Oxyacantha (Hawthorne), CurcumaLonga (Turmeric), Echinacea Species (Purple Coneflower), EleutherococcusSenticosus (Siberian Ginseng), Ephedra Species, Dietary Fish OilConsumption and Fish Oil Supplementation, Genistein, Ginkgo Biloba,Glycyrrhiza (Licorice), Hypericum Perforatum (St. John's Wort),Hydrastis (Goldenseal) and Other Berberine-Containing Plants,Lactobacillus, Lobelia (Indian Tobacco), Melaleuca Alternifolia, MenthaPiperita, NGNA, Panax Ginseng, Pancreatic Enzymes, Piper Mythisticum,Procyanidolic Oligomers, Pygeum Africanum, Quercetin, SarsaparillaSpecies, Serenoa Repens (Saw palmetto, Sabal serrulata), SilybumMarianum (Milk Thistle), Rosemary/Lemon balm, Selenite, TabebuiaAvellanedae (LaPacho), Taraxacum Officinale, Tanacetum Parthenium(Feverfew), Taxol, Uva Ursi (Bearberry), Vaccinium Myrtillus(Blueberry), Valerian Officinalis, Viscum Album (Mistletoe), Vitamin A,Beta-Carotene and Other Carotenoids, and Zingiber Officinale (Ginger).

As used herein, the term “weight loss diet regimen” or related terms, isused broadly to include any type of weight loss plan used by a subject.Examples of weight loss diet regimens include, but are not limited to,Atkins diet, Beverly Hills diet, Cabbage Soup diet, DietSmart.com diet,DietWatch.com diet, Fit For Life diet, Grapefruit diet, Herbalife diet,High Protein diet, Jenny Craig diet, Juice Fasts diet, Kashi GoLeandiet, Low Fat diet, Mayo Clinic diet, Nutrisystem diet, Perricone diet,Pritkin diet, Ready to Eat diet, Revival Soy diet, Richard Simmons diet,Scarsdale diet, Shakes diet, Slim-Fast diet, Somersizing diet, SouthBeach diet, Special K diet, Subway diet, Sugar Busters diet, Thin ForLife diet, Weight Watchers diet, Zone diet, running, swimming,meditation, yoga, hypnosis, clinical therapy, bicycling, walking,hypnosis, rehabilitory training, a dietary plan provided through adietician, and surgical procedures.

As used herein, the terms “subject” and “patient” refer to any animal,such as a mammal like a dog, cat, bird, livestock, and preferably ahuman. Specific examples of “subjects” and “patients” include, but arenot limited to, individuals suffering from viral obesity.

As used herein, the term “physiologically acceptable carrier” refers toany carrier or excipient commonly used with oily pharmaceuticals. Suchcarriers or excipients include, but are not limited to, oils, starch,sucrose and lactose.

As used herein, the term “oral delivery vehicle” refers to any means ofdelivering a pharmaceutical orally, including, but not limited to,capsules, pills, tablets and syrups.

As used herein, the term “food product” refers to any food or feedsuitable for consumption by humans, non-ruminant animals, or ruminantanimals. The “food product” may be a prepared and packaged food (e.g.,mayonnaise, salad dressing, bread, or cheese food) or an animal feed(e.g., extruded and pelleted animal feed or coarse mixed feed).“Prepared food product” means any pre-packaged food approved for humanconsumption.

As used herein, the term “foodstuff” refers to any substance fit forhuman or animal consumption.

As used herein, the term “dietary supplement” refers to a small amountof a compound for supplementation of a human or animal diet packaged insingle or multiple does units. Dietary supplements do not generallyprovide significant amounts of calories but may contain othermicronutrients (e.g., vitamins or minerals).

As used herein, the term “nutritional supplement” refers to acomposition comprising a “dietary supplement” in combination with asource of calories. In some embodiments, nutritional supplements aremeal replacements or supplements (e.g., nutrient or energy bars ornutrient beverages or concentrates).

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the field of nutraceuticals, and inparticular to nutraceuticals comprising n-glycolylneuraminic acid(NGNA). Compositions comprising NGNA find use in inducing physiologicalresponses such as alleviating the symptoms of viral-induced obesity,alleviating the symptoms of colds, preventing the onset of colds,increasing energy and increasing the feeling of well-being in subjects.

A. Treatment and prevention of Viral Infection

Despite many decades of research there are still relatively feweffective anti-viral compounds in comparison to the human disease burdeninflicted by viruses. Coupled with the high mutation rate of certainviruses, which enable anti-viral resistant mutants to arise withalarming speed and frequency, the need for a generic anti-viral agent isas important now as it ever has been.

Influenza has been established as a serious human affliction that cancause localized epidemics and global pandemics of acute respiratoryinfections. Each year the Influenza virus is responsible for 20,000 to40,000 deaths and up to 300,000 hospitalization cases in the UnitedStates (Sandhu & Mossad, 2001). In the pandemic of 1918 it is widelybelieved that in excess of 40 million people died. Although children andyounger adults experience more cases of infection, severe illness ismore common in the elderly, immunocompromised individuals, or those withchronic illnesses such as asthma, diabetes, kidney failure and heartdisease. The annual epidemics run from November to March in the NorthernHemisphere, and from April to September in the Southern Hemisphere (CoxN. J & Subbarao K, 2000).

The two classes of Influenza viruses that are responsible forsignificant human disease are termed Influenza A and Influenza Bviruses. Influenza B viruses are thought to only infect humans whereasInfluenza A viruses are known to infect a wide variety of animalspecies. Both are responsible for seasonal epidemics and, as such, areincluded in the annual influenza vaccines.

Herpes Simplex Virus type 1 (HSV-1) is a member of Herpesviridae, whichcauses infections in humans. HSV-1 has a worldwide distribution with anestimated 60-95% of the adult population infected (Brady, R. C. &Bernstein, D. I., 2004). HSV-1 is frequently associated with oraldisease, normally resulting in the formation of facial lesions. This isan important neurotropic virus which can infect the central nervoussystem (Immergluck et al., 1998). Once access is gained by the virus tothe neurons in sensory ganglia a latent infection can be established(Kramer et al., 2003). Very little is known about the biologicalprocesses involved, but under certain stress conditions reactivation istriggered, typically resulting in the reemergence of the externallesions (Itzhaki & Lin, 1998; Qiu and Abdel-Meguid 1999).

Human Rhinoviruses (HRV) are a member of the Picornaviridae. There havebeen more than 100 different serotypes identified to date. This diversefamily of viruses is thought to be responsible for more than 80% ofupper respiratory tract infections (Heikkinen & Jarvinen, 2003). As thepredominant cause of the common cold, Rhinoviruses are a major humanpathogen both in terms of the disease they cause and the economic impacton society.

Adenoviruses are a group of viruses which cause a wide range ofdiseases. They are known to infect the lining of the eyes, therespiratory tract, the intestines, and the urinary tract and are alsoone of the causative agents of common cold symptoms. The virus can besplit into six sub-groups from A to F based on biological and antigeniccharacteristics (Horwitz, 1996; Shenk 1996). Treatment advice foradenovirus infection is usually bed rest and associated with thetreatment of the symptoms, as at present there is no effective specificantiviral for this virus.

Noroviruses are the most common cause of viral gastroenteritis (Caul etal., 1993; Hedberg & Osterholm, 1993). Consequently, Norovirus issometimes known as the ‘winter vomiting virus’ because it is moreevident during the winter period. The virus is extremely contagious andis found in the vomit and faeces of infected individuals. Currently,Noroviruses cannot be grown in the laboratory by conventional means.However, in the last year some progress has been made on propagationtechniques that involve the use of bioreactors with small intestinalepithelial cells (Straub et al., 2007). However, currently, it isnecessary to use a surrogate virus to test the potential virucidaleffects against Noroviruses. The most established of the surrogateNorovirus viruses is the Feline Calicivirus, which like Norovirus,belongs to the Caliciviridae family of viruses (Slomka et al., 1998;Doultree et al., 1999).

In some preferred embodiments, the present invention provides NGNAcompositions methods of using NGNA compositions for the preventionand/or treatment of viral infections caused by members of the followingfamilies of viruses: Picornaviridae, Caliciviridae, Adenoviridae,Herpesviridae, and Orthomyxoviridae. NGNA is a compound that is foundnaturally in sea cucumbers. NGNA is also found on the surface of allmammalian cells except for humans and chimpanzees and it is thought toact as a viral decoy or scavenger receptor. In some preferredembodiments, the present invention provides NGNA compositions methods ofusing NGNA compositions for the prevention and/or treatment of viralinfections caused by the following viruses: Influenza A, B and C virusesand Asian bird flu viruses (e.g., viruses from the familyOrthomyxoviridae, and in particular Influenza A serotypes H1N1, H2N2,H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3, and H10N7), Herpes SimplexVirus-1 (HSV-1), Rhinoviruses (such as Human Rhinovirus, HRV),Adenoviruses, and Noroviruses. Exemplary formulations are described indetail below. However, in some embodiments, NGNA is formulated as alotion, spray, gel, ointment, powder, aqueous or non-aqueous solutionfor topical, intranasal, intravaginal, intraanal, or sublingualadministration; as a capsule, powder, or tablet for enteraladministration; or as a solution for parenteral administration.

B. Viral Obesity

In the U.S., the prevalence of obesity increased by 30% from 1980 to1990, and this increase appears to be continuing. One etiology of humanobesity is of infectious origin (see, e.g., Dhurandhar, N. (2001) J.Nutr. 131: 2794S-2797S; herein incorporated by reference in itsentirety). Six pathogens are reported to cause obesity in animals.Canine distemper virus was the first virus reported to cause obesity inmice (see e.g., Lyons M., et al., (1982) Science 216:82-85; Bernard A.,et al., (1988) Comp. Biochem. Physiol. 91B:691-696; Bernard A., et al.,(1991) Virology 313:545-551; Bernard A., et al., (1993) J. Neuropath.Exp. Neuro. 52:471-480; McFerran J., et al., (1975) Am. J. Vet. Res.36:527-534; Bernard A., et al., (1999) J. Virology 73(9):7317-7327; eachherein incorporated by reference in their entireties), followed by theavian retrovirus Rous-associated virus-7 shown to cause stunting,obesity and hyperlipidemia in chickens (see e.g., Carter J. K., et al.,(1983) Infect. Immun. 39:410-422; Carter J. K., et al., (1983) Avian Dis27:317-322; each herein incorporated by reference in their entireties).The obesity-promoting effect of Borna disease virus has beendemonstrated in rats (see e.g., Gosztonyi G. & Ludwig H. (1995) CurrentTopics in Microbiol. Immunol. 190:39-73; Fabricant C. G., et al., (1983)Federation Proc 42:2476-2479; each herein incorporated by reference intheir entireties). Scrapie agents were reported to induce obesity inmice and hamsters (see e.g., Kim Y. S., et al., (1987) J. Infect. Dis.August; 156(2):402-405; Carp R. L., et al., (1990) J. Infect. Dis.161(3):462-466; Carp R. L., et al., (1998) J. Gen. Virol. 79(Pt11):2863-2869; Kim Y. S., et al., Proc. Soc. Exp. Biol. Med.189(1):21-27; each herein incorporated by reference in theirentireties). The final two reports were of SMAM-1, an avian adenovirus(see e.g., Dhurandhar N. V., et al., (1990) J. Bombay Vet. College2:131-132; Dhurandhar N. V., et al., (1992) Vet. Microbiol. 31:101-107;Ajinkya S. M. (1985) Final Technical Report, ICAR 1985:13-43 Red andBlue Cross Publishing Bombay, India; each herein incorporated byreference in their entireties), and Ad-36 (see e.g., Dhurandhar N. V.,et al., (2000) Int. J. Obesity 24:989-996; Dhurandhar N. V., et al.,(2001) Int. J. Obesity 25:990-996; Kolesar J. M., et al., (2000) J.Chromatog. B. 744:1-8; herein incorporated by reference in itsentirety), a human adenovirus that caused obesity in animals.Additionally, an association with human obesity is the unique feature ofSMAM-1 (see, e.g., Dhurandhar N. V., et al., (1997) Obesity Res5:464-469; herein incorporated by reference in its entirety) and Ad-36(see, e.g., Vangipuram, S. D., et al., (2004) Obesity Research12:770-777; Dhurandhar, N. V., et al., (2002) J. Nutr. 132:3155-3160;Atkinson R. L., et al., (1998) Int. J. Obesity 22:S57; Dhurandhar N. V.,et al. (1997) FASEB J 3:A230; Atkinson R. L., et al., (2000) Int. J.Obesity suppl 1:S39; Dhurandhar N. V., et al.; (1999) FASEB J13(4):A369; each herein incorporated by reference in their entireties).

U.S. Pat. Nos. 6,127,113 and 6,664,050, each herein incorporated byreference in their entireties, characterize Ad-36 induced viral obesity.In particular, humans who suffer from viral obesity (i.e., obesitycaused by a virus) have, on the average, significantly lowertriglycerides (TG), cholesterol (CHOL) andlow-density-lipoprotein-associated cholesterol (LDL-CHOL) levels thanpersons who suffer from obesity that is not viral. In fact, the averageTG, CHOL and LDL-CHOL levels of persons with viral obesity are withinthe normal ranges for persons who are not obese. Diagnostic andscreening approaches for Ad-36 and Ad-36p are provided. Anti-obesityvaccines, wherein inactivated viral nucleic acid (e.g., Ad-36 nucleicacid) is administered to a subject, are also described.

C. Formulation of NGNA Compositions

NGNA compositions of the present invention may be delivered in anysuitable format. In some embodiments, NGNA is provided from seacucumbers, e.g., an extract of sea cucumbers, or is prepared fromchitin. In some embodiments, the NGNA is preferably about greater than90%, 95%, 99% or 99.9% pure. In some embodiments, the NGNA is HPLCpurified. In some embodiments, NGNA is prepared as described in WO00/38967, incorporated by reference herein its entirety. For example,N-glycolylneuraminic acid can be purchased commercially from, forexample, Sigma Chemical Company, St. Louis, Mo. N-glycolylneuraminicacid also can be synthesized. For example, CMP-N-acetylneuraminic acidhydroxylase can be used to synthesize N-glycolylneurarninic acid as itsCMP-glycoside. See, Schlenzka et al., Glycobiolog, 1994, 4(5):675-683.Non-enzymatic methods of synthesis include, for example, synthesis fromN-acetylneuraminic acid using methanol or hydrochloric acid andbenzylalcohol. Other synthesis methods are described in Choi et al., J.Org. Chem., 1996, 61:8/39 (from mannosamine), Faillard et al., J.Physiol. Chem.' 1965, 344:167 (from glucosamine), U.S. Pat. No.4,774,326 and U.S. Pat. No. 4,774,327, both of which are incorporated byreference herein in their entirety.

In some embodiments, the NGNA compositions are provided in an aqueoussolution, including gels, suitable for use as a spray or mist. In someembodiments, the aqueous NGNA solution is incorporated into a pump-spraycontainer, such as precompression pump, or a device such as a nebulizeror cold mist system, for delivery into the nose, mouth or lungs as afine mist or spray.

In some embodiments, the NGNA compositions of the present inventioncontain a pharmaceutically acceptable excipient which is effective informing a thixotropic suspension of the solid particles of medicamentcomprising the composition, such as those described in U.S. Pat. No.7,122,206. The excipient is preferably present in an amount whichmaintains the particles of medicament suspended in the compositionduring non-use and during spray of the composition into the nasalcavity, and also when the composition is deposited on the mucosalsurfaces of the nasal cavities. In some embodiments, the viscosity ofthe composition at rest is relatively high, for example, about 400 toabout 1000 cp. As the composition is subjected to shear forces, forexample, upon being subjected to forces involved in its being agitatedbefore spraying, the viscosity of the composition decreases (forexample, to about 50 to about 200 cp) and it flows readily through thespray device and exits therefrom in the form of a fine plume whichinfiltrates and deposits on the mucosal surfaces of at least thefollowing parts of the nose: the anterior regions of the nose (frontalnasal cavities); the frontal sinus; the maxillary sinuses; and theturbinates which overlie the conchas of the nasal cavities. Thus, theNGNA compositions comprise a freely flowable liquid, and in sprayedform, a fine mist that finds its way to and deposits on the desiredmucosa. In deposited and relatively unstressed form, the compositionincreases in viscosity and assumes its gel-like form which includesparticles of the medicament suspended therein and which resists beingcleared from the nasal passages by the inherent mucocillary forces thatare present in the nasal cavities.

Any pharmaceutically acceptable material which is capable of maintainingthe solid particles of medicament dispersed substantially uniformly inthe composition and of imparting to the composition desired thixotropicproperties can be used. Such material is referred to as a “suspendingagent”. Examples of suspending agents include carboxmethylcellulose,veegum, tragacanth, bentonite, methylcellulose, and polyethyleneglycols. A preferred suspending agent is a mixture of microcrystallinecellulose and carboxymethylcellulose, the former being presentpreferably in a major amount, most preferably in an amount of about 85to about 95 wt. %, with the latter constituent comprising about 5 toabout 15 wt. % of the mixture.

The amount of suspending agent comprising the composition will varydepending on the particular medicament and amount used, the particularsuspending agent used, the nature and amounts of the other ingredientscomprising the composition, and the particular viscosity values that aredesired. Generally speaking, it is believed that the most widely usedcompositions will comprise about 1 to about 5 wt. % of the suspendingagent.

The NGNA compositions of the present invention includes preferably otheringredients which impart desired properties to the composition. In someembodiments, dispersing or wetting agents are utilized. Any dispersingagent which is effective in wetting the particles and which ispharmaceutically acceptable can be used. Examples of dispersing agentsthat can be used are fatty alcohols, esters, and ethers, including, forexample, those sold under the trademarks Pluronic, Tergitol, Span, andTween. It is preferred to use a hydrophilic, non-ionic surfactant.Excellent results have been achieved utilizing sorbitanmonooleatepolyoxyethylene which is available under the trademarkPolysorbate 80.

In some embodiments, the compositions comprise an anti-oxidant. Examplesof pharmaceutically acceptable anti-oxidants that can be used in thecomposition include ascorbic acid, sodium ascorbate, sodium bisulfite,sodium thiosulfate, 8-hydroxy quinoline, and N-acetyl cysterine. It isrecommended that the composition comprise about 0.001 to about 0.01 wt.% of the anti-oxidant.

Also, for stability purposes, the NGNA compositions should be protectedfrom microbial contamination and growth. Examples of pharmaceuticallyacceptable anti-microbial agents that can be used in the compositioninclude quaternary ammonium compounds, for example, benzalkoniumchloride, benzethonium chloride, cetrimide, and cetylpyridiniumchloride; mercurial agents, for example, phenylmercuric nitrate,phenylmercuric acetate, and thimerosal; alcoholic agents, for example,chlorobutanol, phenylethyl alcohol, and benzyl alcohol; antibacterialesters, for example, esters of para-hydroxybenzoic acid; and otheranti-microbial agents such as chlorhexidine, chlorocresol, andpolymyxin. It is recommended that the composition comprise about 0.001to about 1 wt. % of the anti-microbial agent.

As mentioned above, an aspect of the present invention comprises acomposition which is odorless and which contains a mixture ofstabilizing agents which function as an anti-oxidant and as ananti-microbial agent. The mixture comprises a quaternary ammoniumcompound that has anti-microbial properties and a material which isgenerally recognized as a chelating agent. The use in the composition ofthis combination of materials with the medicament, for example,triamcinolone acetonide, results in a highly stable composition that isresistant to oxidative degradation and to the growth of bacteria and thelike. In preferred form, the mixture comprises benzalkonium chloride anddisodium ethylenediamine tetraacetate.

The odorless composition generally will comprise about 0.004 to about0.02 wt. % of the quaternary ammonium compound and about 0.01 to about0.5 wt. % of the chelating agent. By virtue of the use of theaforementioned mixture of compounds, it is not necessary to include inthe composition a material which is considered an anti-oxidant.

The composition of the present invention includes preferably aniso-osmotic agent which functions to prevent irritation of nasal mucosaby the composition. Dextrose in anhydrous form is a preferrediso-osmotic agent. Examples of other pharmaceutically acceptableiso-osmotic agents which can be used include sodium chloride, dextroseand calcium chloride. It is recommended that the composition comprise upto about 5 wt. % of the iso-osmotic agent.

The pH of the NGNA compositions will vary depending on the particularmedicament used and taking into account biological acceptance and thestability of the medicament. Typically, the pH of the composition willfall within the range of about 4.5 to about 7.5. The preferred pH for acomposition which contains triamcinolone acetonide is about 4.5 to about6, most preferably about 5. Examples of pharmaceutically acceptablematerials which can be used to adjust the pH of the composition includehydrochloric acid and sodium hydroxide.

The NGNA compositions of the present invention can be prepared in anysuitable way. In preferred form, an aqueous suspension of the solidparticles of medicament and dispersing agent is formed and combined withan aqueous suspension which contains the suspending agent. The former ispreferably prepared by adding the medicament to an aqueous solution ofthe dispersing agent and mixing thoroughly. The latter is prepared byacidifying the water (pH about 4.7 to about 5.3) prior to adding thesuspending agent. In particularly preferred form, an aqueous solution ofthe quaternary compound (anti-microbial agent) is added to the aqueoussuspension of medicament, and the other ingredients (for example,iso-osmotic agent, anti-oxidant or chelating agent) are added to thethixotropic suspension. Each of the aforementioned batches ofcomposition is mixed thoroughly before being combined. The preferredmeans of combining the batches of composition is to introduce one of thebatches, preferably the “medicament” batch into the bottom of the otherbatch, for example, by pumping the batch upwardly through the otherbatch. The composition comprising the combined batches is mixedthoroughly. Use of the preferred method of preparation provides anefficient and effective way for formulating a composition that has thesolid particles of medicament substantially uniformly dispersed thereinwhile avoiding problems that are generally associated with thepreparation of water-based pharmaceutical compositions, for example,excessive foaming and non-uniformity of the particle dispersement.

The amount of NGNA applied to each of the nasal passages will varydepending on the nature of the condition being treated and the nature ofthe individual being treated. For guideline purposes, it is suggestedthat that unit dosage applied to one of the nasal cavities compriseabout 5 micrograms to about 100 milligrams NGNA, preferably from about100 micrograms too about 1 milligram NGNA. Use of the preferred form ofthe composition of the present invention provides the advantage that thecomposition can be applied effectively once daily.

Accordingly, the present invention provides an article of manufacturecomprising a spray bottle having an NGNA solution or powder therein fordelivery into a body cavity such as the nose. The spray bottle maypreferably comprise a pump system for expelling the NGNA compositionfrom the bottle, such as a compression mump, spray pump orprecompression pump.

In some embodiments, the present invention provides an article ofmanufacture that is a device that can be worn over a body cavity such asthe mouth or nose of an individual. In some embodiments, the device ismask, such as a surgical mask. In preferred embodiments, the maskcomprises a solid support or matrix, such as a polymer matrix or a wovenfabric matrix, into which an NGNA composition is incorporated. In someembodiments, the NGNA composition is spray coated onto the matrix as anaqueous solution, gel or powder. In some embodiments, when a breath istaken through the mask, viruses are inactivated as they contact themask. In some embodiments, the matrix is coated with an NGNA compositionto provide from about 0.01 microgram to about 100 milligram NGNA per cm²of the matrix, preferably from about 1 microgram to about 1 milligramNGNA per cm².

In some embodiments, NGNA is formulated for oral delivery. Theingredients of the supplements of this invention are preferablycontained in acceptable excipients and/or carriers for oral consumption.The actual form of the carrier, and thus, the supplement itself, is notcritical. The carrier may be a liquid, gel, gelcap, capsule, powder,solid tablet (coated or non-coated), tea, or the like. The supplement ispreferably in the form of a tablet or capsule and most preferably in theform of a hard gelatin capsule. Suitable excipient and/or carriersinclude maltodextrin, calcium carbonate, dicalcium phosphate, tricalciumphosphate, microcrystalline cellulose, dextrose, rice flour, magnesiumstearate, stearic acid, croscarmellose sodium, sodium starch glycolate,crospovidone, sucrose, vegetable gums, lactose, methylcellulose,povidone, carboxymethylcellulose, corn starch, and the like (includingmixtures thereof). Preferred carriers include calcium carbonate,magnesium stearate, maltodextrin, and mixtures thereof. The variousingredients and the excipient and/or carrier are mixed and formed intothe desired form using conventional techniques. The tablet or capsule ofthe present invention may be coated with an enteric coating thatdissolves at a pH of about 6.0 to 7.0. A suitable enteric coating thatdissolves in the small intestine but not in the stomach is celluloseacetate phthalate. Further details on techniques for formulation for andadministration may be found in the latest edition of Remington'sPharmaceutical Sciences (Maack Publishing Co., Easton, Pa.).

In some embodiments, the oral delivery vehicles described above areformulated so as to provide a daily dose of between 0.1 g and 10.0 g ofNGNA, preferably between 0.5 and 2.0 g of NGNA, and even more preferablyapproximately 1.0 g of NGNA. In some embodiments, an effective amount ofNGNA is the amount needed to inhibit the growth and proliferation of avirus of interest or to inactivate the virus. In some embodiments, theeffective amount is the amount of NGNA sufficient to provide aconcentration of NGNA of from about 1 nanomolar to 10 micromolar at asite of interest, such as the circulating bloodstream or in a bodycavity such as the nasal passages or sinuses.

In some embodiments, NGNA is provided in a fluid that can be used foratmospheric treatment, such as by a mist. In some embodiments, thepresent invention provides a device comprising a reservoir, a pump, anda nozzle, wherein the reservoir comprises a fluid comprising NGNA thatcan be expelled via the pump through the nozzle to provide a mistcomprising NGNA. In some embodiments, the device is a humidifier, whilein other embodiments, the device is an automated mist dispenser. In someembodiments, the NGNA is provided as an aerosol spray in an appropriateaerosol spray dispensing device. Accordingly, in some embodiments, thepresent invention provides a device or composition comprising a NGNA andan aerosol propellant. Propellants include, but are not limited to,mixtures of volatile hydrocarbons, typically propane, n-butane andisobutene, dimethyl ether (DME), methyl ethyl ether, nitrous oxide,carbon dioxide and hydrofluoroalkanes (HFA): either HFA 134a(1,1,1,2,-tetrafluoroethane) or HFA 227(1,1,1,2,3,3,3-heptafluoropropane) or combinations of the two.Typically, the NGNA fluid will be miscible with the propellant. The NGNAfluid may preferably be formulated to be an aerosol mist, foaming gel,cream or lotion.

Modern aerosol spray products have three major parts; the can, the valveand the actuator or button. The can is most commonly lacquered tinplate(steel with a layer of tin) and may be made of 2 or 3 pieces of metalcrimped together. Aluminum cans are also common and are generally usedfor more expensive products. The valve is crimped to the rig of the can,the design of this component is important in determining the spray rate.The actuator is depressed by the user to open the valve; the shape andsize of the nozzle in the actuator controls the spread of the aerosolspray.

In some embodiments, the devices of the present invention comprise apiston barrier system. Packaging that uses a piston barrier system isoften used for highly viscous products such as post-foaming gels, thickcreams and lotions. The main benefit of the piston barrier system isthat is assures separation of the product from the propellant,maintaining the purity and integrity of the formulation throughout itsconsumer lifespan. The piston barrier system also provides a controlledand uniform product discharge rate with minimal product retention and iseconomical.

In some embodiments, the devices of the present invention comprise abag-in-can system (or BOV “bag on valve”). This system separates theproduct from the pressurizing agent with a hermetically-sealed,multi-layered laminated pouch, which maintains complete formulationintegrity so only pure product is dispensed. In this embodiment, theNGNA fluid is provided in the bag. Among its many benefits, thebag-in-can system extends a product's shelf life. Other advantagesinclude all-attitude (360-degree) dispensing, quiet and non-chillingdischarge.

These devices find use in setting where an antiviral spray, foam, gel orother fluid is needed. For example, the devices find use in dispensingan antiviral mist to treat a desired environment or space, such as aroom in a building such as an office, kitchen, cubicle or lavatory, or avehicle such a train car, airplane, bus, or taxi, van or car, or animalconfinement facilities such as barns, abattoirs, poultry houses, etc.The devices can also be used to dispense NGNA containing fluids on asurface such as a countertop, floor, shower, etc. In some embodiments,the devices are used to inject the NGNA-containing mist into theventilation system of a building, barn, or vehicle.

In still further embodiments, the present invention provides NGNAcompositions that comprise NGNA in a solution, such as a normal salinesolution, that can be applied to the eye. Accordingly, in someembodiments, the present invention provides an article of manufacturecomprising a container equipped with a nozzle to provide drops of anNGNA solution to the eye. It is contemplated that in addition to effectsin the eye and surrounding tissue, NGNA administration via the eye andtear channel will directly access the sinuses and associated cavities.Thus, administration of NGNA and/or additional antimicrobials orantivirals in eyedrops is an effective method of administration of NGNAor other compounds for the treatment of eye infections, sinusinfections, and systemic treatment via the eye and tear channel, andsinuses, mucus internal surfaces for effective adsorption providinglocal and systemic treatments.

In other embodiments, the NGNA is provided as a powder or liquidsuitable for adding by the consumer to a food or beverage. For example,in some embodiments, the dietary supplement can be administered to anindividual in the form of a powder, for instance to be used by mixinginto a beverage, or by stirring into a semi-solid food such as apudding, topping, sauce, puree, cooked cereal, or salad dressing, forinstance, or by otherwise adding to a food.

In some embodiments, the NGNA is provided in water that is supplied tofarm animals, such as poultry, cattle, swine, sheep and the like, orused in water used in fisheries. In other embodiments, NGNA is providedin tap water or water bottles water for human use.

The NGNA compositions may comprise one or more inert ingredients,especially if it is desirable to limit the number of calories added tothe diet by the dietary supplement. For example, the dietary supplementof the present invention may also contain optional ingredientsincluding, for example, herbs, vitamins, minerals, enhancers, colorants,sweeteners, flavorants, inert ingredients, and the like. For example,the compositions of the present invention may contain one or more of thefollowing: asorbates (ascorbic acid, mineral ascorbate salts, rose hips,acerola, and the like), dehydroepiandosterone (DHEA), Fo-Ti or Ho Shu Wu(herb common to traditional Asian treatments), Cat's Claw (ancientherbal ingredient), green tea (polyphenols), inositol, kelp, dulse,bioflavinoids, maltodextrin, nettles, niacin, niacinamide, rosemary,selenium, silica (silicon dioxide, silica gel, horsetail, shavegrass,and the like), spirulina, zinc, and the like. Such optional ingredientsmay be either naturally occurring or concentrated forms.

In some embodiments, the compositions further comprise vitamins andminerals including, but not limited to, calcium phosphate or acetate,tribasic; potassium phosphate, dibasic; magnesium sulfate or oxide; salt(sodium chloride); potassium chloride or acetate; ascorbic acid; ferricorthophosphate; niacinamide; zinc sulfate or oxide; calciumpantothenate; copper gluconate; riboflavin; beta-carotene; pyridoxinehydrochloride; thiamin mononitrate; folic acid; biotin; chromiumchloride or picolonate; potassium iodide; sodium selenate; sodiummolybdate; phylloquinone; vitamin D₃; cyanocobalamin; sodium selenite;copper sulfate; vitamin A; vitamin C; inositol; potassium iodide.Suitable dosages for vitamins and minerals may be obtained, for example,by consulting the U.S. RDA guidelines.

D. Dietary Supplements

The present invention provides dietary supplements comprisingnutraceutical agents, preferably NGNA either by itself or in combinationwith one or more additional nutraceuticals agents. Nutraceutical agentsare natural, bioactive chemical compounds that have health promoting,disease preventing or medicinal properties. Examples of nutraceuticalsinclude, but are not limited to, Allium Cepa, Allium Sativum, Aloe Vera,Angelica Species, Naturally Occurring Antioxidants, Aspergillus OryzaeEnzyme Therapy, barley grass, Bromelain, Carnitine, Carotenoids andFlavonoids, Catechin, Centella Asiatica (Gotu kola), Coenzyme Q10,Chinese Prepared Medicines, Coleus Forskohlii, Commiphora Mukul,Conjugated Linoleic Acids (CLAs), Crataegus Oxyacantha (Hawthorne),Curcuma Longa (Turmeric), Echinacea Species (Purple Coneflower),Eleutherococcus Senticosus (Siberian Ginseng), Ephedra Species, DietaryFish Oil Consumption and Fish Oil Supplementation, Genistein, GinkgoBiloba, Glycyrrhiza (Licorice), Hypericum Perforatum (St. John's Wort),Hydrastis (Goldenseal) and Other Berberine-Containing Plants,Lactobacillus, Lobelia (Indian Tobacco), Melaleuca Alternifolia,Menaquinone, Mentha Piperita, n-glycolylneuraminic acid (NGNA), PanaxGinseng, Pancreatic Enzymes, Piper Mythisticum, Procyanidolic Oligomers,Pygeum Africanum, Quercetin, Sarsaparilla Species, Serenoa Repens (Sawpalmetto, Sabal serrulata), Silybum Marianum (Milk Thistle),Rosemary/Lemon balm, Selenite, Tabebuia Avellanedae (LaPacho), TaraxacumOfficinale, Tanacetum Parthenium (Feverfew), Taxol, Uva Ursi(Bearberry), Vaccinium Myrtillus (Blueberry), Valerian Officinalis,Viscum Album (Mistletoe), Vitamin A, Beta-Carotene and OtherCarotenoids, and Zingiber Officinale (Ginger).

Several nutraceutical agents are used in treating viral disorders (e.g.,Genistein (in soy/red clover), rosemary/lemon balm, selenite, barleygrass, lauric acid, Phyllanthus amarus/niruri (see, e.g., Nicolson, G.(1998) J. Medicine 1:123-128; herein incorporated by reference in itsentirety). Additional anti viral nutraceutical agents include, but arenot limited to, catechins, flavenoids, Echinacea, cascara, and NGNA.

In preferred embodiments, the present invention provides compositionscomprising dietary supplements (e.g., NGNA) for inducing physiologicalresponses such as alleviating the symptoms of viral-induced obesity,alleviating or treating the symptoms of colds, preventing the onset ofcolds, increasing energy and increasing the feeling of well-being insubjects. Such compositions may contain, for example, between 0.1 g and10.0 g of dietary supplements (e.g., NGNA), preferably between 0.5 g and2.0 g of dietary supplements (e.g., NGNA), and even more preferably,approximately 1.0 g of dietary supplements (e.g., NGNA). Furthermore,dietary supplements (e.g., NGNA) are preferably provided in an amountsufficient to induce the physiological response desired (e.g.,alleviation of the symptoms of viral-induced obesity, alleviation of thesymptoms of colds, prevention of the onset of colds, increasing energyand increasing the feeling of well-being in subjects). In someembodiments, the compositions are provided for use in inducing one ofthe foregoing responses, while in other embodiments, the compositionsare provided for use in inducing two or more of the foregoing responses.

The present invention further provides methods for treating thephysiological conditions discussed above (e.g., conditions such asobesity, periodic weight gain, lack of energy, mild depression, colds,etc.). In preferred embodiments, dietary supplements are used intreating viral induced obesity. In other preferred embodiments, dietarysupplements are used in treating infection caused by a virus that causesviral obesity (e.g., Ad-36). In other preferred embodiments, dietarysupplements are used in preventing viral related obesity throughtargeting of viruses that cause viral obesity (e.g., Ad-36, Ad-36p,SMAM-1). In some embodiments, the compositions are provided for use intreating one of the foregoing conditions, while in other embodiments,the compositions are provided for use in treating two or more of theforegoing conditions.

The dietary supplements of the present invention are further used inconjunction with a weight loss diet regimen. The present invention isnot limited to a particular kind of weight loss diet regimen (e.g.,exercise, reduced calorie intake, etc.). In preferred embodiments, theweight loss diet regimen is a dietary plan (e.g., Atkins diet, BeverlyHills diet, Cabbage Soup diet, DietSmart.com diet, DietWatch.com diet,Fit For Life diet, Grapefruit diet, Herbalife diet, High Protein diet,Jenny Craig diet, Juice Fasts diet, Kashi GoLean diet, Low Fat diet,Mayo Clinic diet, Nutrisystem diet, Perricone diet, Pritkin diet, Readyto Eat diet, Revival Soy diet, Richard Simmons diet, Scarsdale diet,Shakes diet, Slim-Fast diet, Somersizing diet, South Beach diet, SpecialK diet, Subway diet, Sugar Busters diet, Thin For Life diet, WeightWatchers diet, and Zone diet. In still other preferred embodiments, theweight loss diet regimen is an exercise plan (e.g., running, swimming,meditation, yoga, hypnosis, clinical therapy, bicycling, walking, etc.).In still other preferred embodiments, the weight loss diet regimen is aclinically assisted plan (e.g., hypnosis, rehabilitory training, adietary plan provided through a dietician, surgical procedures, etc.).

The dietary supplements of the present invention may further beadministered in any form (e.g., pill, food product, etc.). In preferredembodiments, the dietary supplements are provided as a beverage, bar,powder, pill, or shake (e.g., a nutritional supplement as described inmore detail below).

The dietary supplements of the present invention may be taken one ormore times daily. Preferably, the dietary supplement is administeredorally one to two times daily. Frequency of administration will, ofcourse, depend on the dose per unit (capsule or tablet) and the desiredlevel of ingestion. Dose levels/unit can be adjusted to provide therecommended levels of ingredients per day (e.g., approximately 1 g of anutraceutical agent) in a reasonable number of units (e.g., two capsulesor tablets taken twice a day). In preferred embodiments, the doses addup each day to the daily intake of each ingredient. In preferredembodiments, the dietary supplements are taken with meals or beforemeals. In other embodiments, the dietary supplements are not taken withmeals. In preferred embodiments, a dietary supplement increases satietyand results in a decrease in caloric intake and subsequent weight loss.In particularly preferred embodiments, a dietary supplement regulatesviruses (e.g., adenoviruses).

E. Nutritional Supplements

In other embodiments, the present invention provides nutritionalsupplements (e.g., energy bars or meal replacement bars or beverages)comprising NGNA. The nutritional supplement may serve as meal or snackreplacement and generally provide nutrient calories. Preferably, thenutritional supplements provide carbohydrates, proteins, and fats inbalanced amounts. The nutritional supplement can further comprisecarbohydrate, simple, medium chain length, or polysaccharides, or acombination thereof. A simple sugar can be chosen for desirableorganoleptic properties. Uncooked cornstarch is one example of a complexcarbohydrate. If it is desired that it should maintain its highmolecular weight structure, it should be included only in foodformulations or portions thereof which are not cooked or heat processedsince the heat will break down the complex carbohydrate into simplecarbohydrates, wherein simple carbohydrates are mono- or disaccharides.The nutritional supplement contains, in one embodiment, combinations ofsources of carbohydrate of three levels of chain length (simple, mediumand complex; e.g., sucrose, maltodextrins, and uncooked cornstarch).

Sources of protein to be incorporated into the nutritional supplement ofthe invention can be any suitable protein utilized in nutritionalformulations and can include whey protein, whey protein concentrate,whey powder, egg, soy flour, soy milk soy protein, soy protein isolate,caseinate (e.g., sodium caseinate, sodium calcium caseinate, calciumcaseinate, potassium caseinate), animal and vegetable protein andmixtures thereof. When choosing a protein source, the biological valueof the protein should be considered first, with the highest biologicalvalues being found in caseinate, whey, lactalbumin, egg albumin andwhole egg proteins. In a preferred embodiment, the protein is acombination of whey protein concentrate and calcium caseinate. Theseproteins have high biological value; that is, they have a highproportion of the essential amino acids. See Modern Nutrition in Healthand Disease, eighth edition, Lea & Febiger, publishers, 1986, especiallyVolume 1, pages 30-32.

The nutritional supplement can also contain other ingredients, such asone or a combination of other vitamins, minerals, antioxidants, fiberand other dietary supplements (e.g., protein, amino acids, choline,lecithin, omega-3 fatty acids). Selection of one or several of theseingredients is a matter of formulation, design, consumer preference andend-user. The amounts of these ingredients added to the dietarysupplements of this invention are readily known to the skilled artisan.Guidance to such amounts can be provided by the U.S. RDA doses forchildren and adults. Further vitamins and minerals that can be addedinclude, but are not limited to, calcium phosphate or acetate, tribasic;potassium phosphate, dibasic; magnesium sulfate or oxide; salt (sodiumchloride); potassium chloride or acetate; ascorbic acid; ferricorthophosphate; niacinamide; zinc sulfate or oxide; calciumpantothenate; copper gluconate; riboflavin; beta-carotene; pyridoxinehydrochloride; thiamin mononitrate; folic acid; biotin; chromiumchloride or picolonate; potassium iodide; sodium selenate; sodiummolybdate; phylloquinone; vitamin D₃; cyanocobalamin; sodium selenite;copper sulfate; vitamin A; vitamin C; inositol; potassium iodide.

Flavors, coloring agents, spices, nuts and the like can be incorporatedinto the product. Flavorings can be in the form of flavored extracts,volatile oils, chocolate flavorings, peanut butter flavoring, cookiecrumbs, crisp rice, vanilla or any commercially available flavoring.Examples of useful flavoring include, but are not limited to, pure aniseextract, imitation banana extract, imitation cherry extract, chocolateextract, pure lemon extract, pure orange extract, pure peppermintextract, imitation pineapple extract, imitation rum extract, imitationstrawberry extract, or pure vanilla extract; or volatile oils, such asbalm oil, bay oil, bergamot oil, cedarwood oil, walnut oil, cherry oil,cinnamon oil, clove oil, or peppermint oil; peanut butter, chocolateflavoring, vanilla cookie crumb, butterscotch or toffee. In oneembodiment, the dietary supplement contains cocoa or chocolate.

Emulsifiers may be added for stability of the final product. Examples ofsuitable emulsifiers include, but are not limited to, lecithin (e.g.,from egg or soy), and/or mono- and di-glycerides. Other emulsifiers arereadily apparent to the skilled artisan and selection of suitableemulsifier(s) will depend, in part, upon the formulation and finalproduct.

Preservatives may also be added to the nutritional supplement to extendproduct shelf life. Preferably, preservatives such as potassium sorbate,sodium sorbate, potassium benzoate, sodium benzoate or calcium disodiumEDTA are used.

In addition to the carbohydrates described above, the nutritionalsupplement can contain natural or artificial (preferably low calorie)sweeteners, e.g., saccharides, cyclamates, aspartamine, aspartame,acesulfame K, and/or sorbitol. Such artificial sweeteners can bedesirable if the nutritional supplement is intended to be consumed by anoverweight or obese individual, or an individual with type II diabeteswho is prone to hyperglycemia.

The nutritional supplement can be provided in a variety of forms, and bya variety of production methods. In a preferred embodiment, tomanufacture a food bar, the liquid ingredients are cooked; the dryingredients are added with the liquid ingredients in a mixer and mixeduntil the dough phase is reached; the dough is put into an extruder, andextruded; the extruded dough is cut into appropriate lengths; and theproduct is cooled. The bars may contain other nutrients and fillers toenhance taste, in addition to the ingredients specifically listedherein.

Servings of the nutritional supplement preferably contain between 0.1 gand 10.0 g of NGNA, preferably between 0.5 and 2.0 g NGNA, and even morepreferably approximately 1.0 g NGNA. It is understood by those of skillin the art that other ingredients can be added to those describedherein, for example, fillers, emulsifiers, preservatives, etc. for theprocessing or manufacture of a nutritional supplement.

F. Food Products

In still further embodiments, the present invention provides functionalfoods, including food products, prepared food products, or foodstuffscomprising nutraceutical agents. For example, in some embodiments,beverages and solid or semi-solid foods comprising nutraceutical agentsare provided. These forms can include, but are not limited to, beverages(e.g., soft drinks, milk and other dairy drinks, and diet drinks), bakedgoods, puddings, dairy products, confections, snack foods, or frozenconfections or novelties (e.g., ice cream, milk shakes), prepared frozenmeals, candy, snack products (e.g., chips), soups, spreads, sauces,salad dressings, prepared meat products, cheese, yogurt and any otherfat or oil containing foods, and food ingredients (e.g., wheat flour).

Servings of the food product preferably contain between 0.1 g and 10.0 gof NGNA, preferably between 0.5 and 2.0 g of NGNA, and even morepreferably approximately 1.0 g of NGNA.

G. Administration to Animals

In some embodiments, the present invention contemplates administrationof NGNA to animals. Preferred animals include, but are not limited,poultry, fish, companion animals such as dogs and cats, horses, swine,and ruminants such as cattle and sheep. The NGNA may be administered forany of the purposes described above, e.g., prevention of viralinfection, treatment of viral infections, etc. In some preferredembodiments, the present invention provides NGNA compositions methods ofusing NGNA compositions for use in animals for the prevention and/ortreatment of viral infections caused by members of the followingfamilies of viruses: Picornaviridae, Caliciviridae, Adenoviridae,Herpesviridae, and Orthomyxoviridae. Exemplary formulations aredescribed in detail above. However, in some embodiments, NGNA isformulated as a lotion, spray, gel, ointment, powder, aqueous ornon-aqueous solution for topical, intranasal, intravaginal, intraanal,or sublingual administration; as a capsule, powder, or tablet forenteral administration; or as a solution for parenteral administration.In some embodiments, the NGNA is preferably provided as a component of apowder that can be dissolved in a water supply for animals, or used tobathe animals. In some embodiments, the NGNA is provided as a liquidthat can be aerosolized or misted into a barn or other enclosurecontaining animals. In other embodiments, the NGNA is supplied as asolution that can be injected into a water system designed for animals.

In some embodiments, the present invention provides animal feedscomprising NGNA. The animal feeds are preferably formulated as in knownin the art. Exemplary animal feeds include diets formulated for poultry,fish, companion animals such as dogs and cats, horses, swine, andruminants such as cattle and sheep. In some embodiments, the NGNA isprovided as a supplement that can be incorporated into animal feeds asthey are being mixed or manufactured. In preferred embodiments, thefeeds are formulated so as to provide a daily dose of between 0.1 g and10.0 g of NGNA, preferably between 0.5 and 2.0 g of NGNA, and even morepreferably approximately 1.0 g of NGNA.

EXAMPLES Example 1

A sports drink containing NGNA was formulated for use by humans. TheNGNA was made from glucosamine residues derived from crab chitin. Thefinished drink (per 500 ml bottle) contained: pure water; 1%Glycerol/(0, 1-10%); lemon flavor; potassium sorbate/sodium benzoate asstabilizers; and 2 mg of 99% pure NGNA.

Example 2

The sports drink described in Example 1 was tested in a group of 18subjects. The subjects were supplied one 500 ml bottle of the sportdrink containing 2 mg NGNA. The drink was administered during exerciseor during the day when not exercising over an eight week period. Thesubjects worked out an average of about four times per week. Thefollowing measurements were taken during the study: fett/fat %,vekt/weight, overarm/upper arm in cm, midje/waist in cm, lår /thighs incm. These results are summarized in FIG. 1 b. As can be seen, there wasa steady decrease in weight during the study (FIG. 1 a). The subjectswere also asked to evaluate other physiological effects of the drink.The subjects were asked to rank the impact of the drink on a scale of 0to 9 (with 9 being a positive impact) with respect to the followingquestions:

a. How was your workout today

b. How do you feel?

c. Do you have a cold 0, or are you healthy 9?

d. Do you feel the drink helps you?

e. How is workout energy and joy?

f. Do you like the sport drink?

The results are presented in FIGS. 2 a-f, respectively. As can be seen,there were positive responses for well-being, colds, and energy/workoutjoy.

Example 3

This example describes the in vitro inhibition of various viruses bycompositions comprising NGNA.

Influenza

The Influenza B virus that will be used for this study will beB/Malaysia.

-   -   Source: Retroscreen Virology Ltd. Virus B/Malaysia/2506/2004    -   Physical form: Virus in liquid medium    -   Expiry date: 4 hours after dilution    -   Storage conditions: On ice    -   Safety precautions: Biosafety Level 2 containment conditions

The cell line to be used in Phase A is MDCK cells. The Growth Media forthe MDCK cell line is DMEM, 2 mM L-Glutamine, 10% FBS (foetal bovineserum) and 2 mM Hepes buffer. The Infection Media for the MDCK cell lineis DMEM, 2 mM L-Glutamine, 2 mM Hepes buffer and 1.5 mM Trypsin (TPCKtreated).

In the virucidal assay the Influenza B virus titrations will beincubated for 2 to 3 days before the efficacy of the test article on thevirus will be determined by HA (Haemagglutination Assay). This assayutilizes the ability of influenza virus to bind red blood cells todetermine the presence or absence of virus in a given well on thetitration plate. The method for this procedure is as follows:

-   -   After the 3 days incubation period of the virus titrations from        the virucidal assay (see section 8.1.1.3), 50 μL of the        supernatant from each well on the 96-well titration plate will        be transferred to the corresponding well on a fresh v-bottomed        96-well assay plate.    -   50 μL of 0.5% turkey red blood cells in PBS (v/v) will then be        added to each well on the v-bottomed assay plate to give a total        volume of 100 mL.    -   The virus/red blood cell suspensions will be mixed by placing        the plates on a plate shaker for approximately 30 seconds at        200-400 rpm.    -   The plates will then be incubated at room temperature for        approximately 30 minutes before being read visually for the        presence of virus in each well.    -   Each well will be scored positive or negative. Briefly,        haemagglutination of the red blood cells will indicate a        positive result and sedimentation of the red blood cells into        the apex of v-shaped well will indicate a negative result.

Herpes Simplex Type 1 (HSV-1)

The Herpes Simplex virus that will be used for this study is HSV-1.

-   -   Source: Retroscreen Virology Ltd. Virus HSV-1, SC-16    -   Physical form: Virus in liquid medium    -   Expiry date: 4 hours from dilution    -   Storage conditions: On ice    -   Safety precautions: Biosafety Level 2 containment conditions

The cell line to be used in Phase B is Vero cells. The Growth Media forthe Vero cell line is DMEM, 2 mM L-Glutamine, 10% FBS (foetal bovineserum) and 2 mM Hepes buffer. The Infection Media is the same as theGrowth Media. In the virucidal assay HSV-1 titrations will be incubatedfor 3 days before the efficacy of the test article on the virus can bedetermined by CPE.

Rhinovirus

The Rhinovirus that will be used for this study is HRV-16

-   -   Source: Retroscreen Virology Ltd. Virus HRV-16    -   Physical form: Virus in liquid medium    -   Expiry date: 4 hours after dilution    -   Storage conditions: On ice    -   Safety precautions: Biosafety Level 2 containment conditions

The cell line to be used in Phase C is Wi-38 cells. The Growth Media forthe Wi-38 cell line is DMEM, 2 mM L-Glutamine, 10% FBS (foetal bovineserum). The Infection Media for the Wi-38 cell line is DMEM, 2 mML-Glutamine, 2% FBS (foetal bovine serum) and 10 mM MgCl₂.

In the virucidal assay, Rhinovirus titrations will be incubated for 4-7days before the efficacy of the test article on the virus can bedetermined by CPE.

Adenovirus

The Adenovirus that will be used for this study is AD-5

-   -   Source: Retroscreen Virology Ltd. Virus AD-5    -   Physical form: Virus in liquid medium    -   Expiry date: 4 hours from dilution    -   Storage conditions: On ice    -   Safety precautions: Biosafety Level 2 containment conditions

The cell line to be used in Phase D is MRC-5 cells. The Growth Media forthe MRC-5 cell line is EMEM, 2 mM L-Glutamine, 10% FBS (foetal bovineserum) and 1× NEAA (non-essential amino acids). The Infection Media forthe MRC-5 cell line is EMEM, 2 mM L-Glutamine, 2% FBS (foetal bovineserum) and 10 mM MgCl₂. In the virucidal assay Adenovirus titrationswill be incubated for 7-10 days before the efficacy of the test articleon the virus can be determined by CPE.

Norovirus Surrogate Virus

The Norovirus-surrogate virus that will be used for this study is FelineCalicivirus (FCV).

-   -   Source: Retroscreen Virology Ltd. Virus Feline Calicivirus (FCV)        (substitute for norovirus)    -   Physical form: Virus in liquid medium    -   Expiry date: 4 hours from diluting    -   Storage conditions: On ice    -   Safety precautions: Biosafety Level 2 containment conditions

The cell line to be used in Phase E is CRFK (Crandell-Reese felinekidney) cells. The Growth Media for the CRFK cell line is EMEM, 2 mML-Glutamine, 10% FBS (foetal bovine serum). The Infection Media for theCRFK cell line is EMEM, 2 mM L-Glutamine, 2% FBS foetal bovine serum).In the virucidal assay FCV titrations will be incubated for 3 to 5 daysbefore the efficacy of the test article on the virus can be determinedby CPE.

Methods

N-Glycolylneuraminic acid (test article) will be tested for the abilityto inhibit the viruses identified above. The physical form of NGNA is aHPLC purified dry powder, Molecular weight 325.268. In the virucidalassay 9 volumes of test article is added to one volume of virus.Therefore, when the virus is mixed with the test article the resultingfinal concentration of the test article will be at 90% (v/v). The testarticle comes in the form of an off white powder, this will be initiallydissolved in dH2O to generate a stock solution before workingconcentrations are made by subsequent dilution in Infection media.

Controls.

The controls to be utilized in the virucidal assay are:

-   -   Cell only control—Cells not infected with virus. This will be a        negative control for vCPE (viral cytopathic effect) and is also        an indicator of cell quality.    -   Virus only control—Cells infected with cell infection media that        has been treated with virus in the same way as the test        articles. This will be a positive control for vCPE.    -   Antiviral control—Cells infected with virus pre-treated with        citrate buffer solution, 0.09 M, pH 3.5. This will be a positive        control for the test articles.

Cytotoxicity Testing.

The test article will be assessed as follows:

-   -   A 100 μM solution of the compound SCN-KI-N5G will be made using        the Infection media relevant to each cell line used in this        study for the viruses being tested (see details for separate        phases in section 8.1). Each cell line in use will be tested.    -   Cells will be seeded into a 96-well plate the day prior to use        in the assay.    -   All the media will be removed from the first row, and 111 mL of        either the test (100 μM SCN-KI-N5G) or control articles will be        added in quadruplicate to the wells.    -   The test/control articles will be titrated in a 10-fold dilution        series for a further 10 rows.    -   The last row of wells will be used as the cell only control.    -   The cells will be then incubated for 4 hours at 37oC. with 5%        CO2.    -   After the incubation period, each well will be checked for tCPE.    -   The tCPE data will be used to determine the highest        concentration of test article that can be used in the virucidal        assay before cytotoxic effects of the test article on the cells        would interfere with the interpretation of the virucidal assay        data.    -   The results of the cytotoxicity assay will be discussed with the        sponsor prior to commencing the virucidal assay, such that an        agreement is reached on what concentrations to use in the        virucidal assay.

Virucidal Testing.

The virucidal assay will be performed as described below. A single timepoint of 10 minutes will be used for each of the 3 differentconcentrations of test article to be tested.

Testing Protocol.

-   -   Add 360M1 of test article at the required test concentration to        40M1 of virus (Total volume=400 mL)    -   Vortex to mix and incubate at room temperature for 10 minutes    -   Terminate the reaction by dilution with the addition of 3.6 mL        infection media (Total volume=4 mL)    -   Inoculate the terminated reaction onto the cell monolayer in        quadruplicate and titrate residual virus by 10-fold serial        dilution    -   Incubate the cells for 2-7 days (as required for the specific        phase, see section 8.1) at 37° C., 5% CO₂.    -   Confirm the residual virus titer using vCPE and/or        Haemagglutination Assay.

Results.

Sample Control titer virus titer (log₁₀ (log₁₀ Virus Cell TCID₅₀/TCID₅₀/ reduction Virus Line Sample ml ml (%) Influenza B MDCK NGNA 10mM 2.50 4.50 99.00 Influenza B MDCK NGNA 10 μM 4.50 4.50 0.00 InfluenzaB MDCK NGNA 10 nM 4.50 4.50 0.00 Influenza B MDCK Citrate buffer 2.504.50 99.00 (+control) HSV-1 Vero NGNA 10 mM 2.50 6.25 99.98 HSV-1 VeroNGNA 10 μM 6.25 6.25 0.00 HSV-1 Vero NGNA 10 nM 7.00 6.25 0.00 HSV-1Vero Citrate buffer 2.50 6.25 99.98 (+control) HRV Wi-38 NGNA 10 mM 2.504.75 99.44 HRV Wi-38 NGNA 10 μM 3.50 4.50 90.00 HRV Wi-38 NGNA 10 nM3.50 4.50 90.00 HRV Wi-38 Citrate buffer 2.50 4.75 99.44 (+control) FCVCRFK NGNA 10 mM 4.50 5.25 82.21 FCV CRFK NGNA 10 μM 5.50 5.50 0.00 FCVCRFK NGNA 10 nM 5.50 5.50 0.00 FCV CRFK Citrate buffer 4.50 5.25 82.11(+control)

Example 4

The overall purpose of the study was to determine whether the N5Gnutritional supplement has the ability to have significant impact onclinical disease outcomes caused by influenza virus in a mouse animalmodel.

The experimental design included adherence to a strict feeding schedule,which included 30 days of acclimation of mice to a defined base feed(AlN-93M) provided by Purina TestDiet, Inc. Control animals remainedcontinuously fed with the defined base feed, while the experimentalmouse group was fed with the base feed supplemented with 0.35% of N5G.

Experimental and control mouse groups were infected intranasally with107.5 TCID50 influenza A (H1N1) with each mouse receiving 5iL of viralinoculum per nostril. The experimental parameters followed during thestudy were: 1) mouse weight changes; 2) food intake changes; 3) clinicalsigns of disease; 4) lung tissue pathology; 5) lung consolidationscores; 6) changes in viral loads.

Overall, the experimental design was executed without any majorobstacles. As expected, there was minor occasional wasting of feed insome cages (mice chewed on feed which was dropped on the bottom of thecage). In every instance noted on daily observations, the residual feedwas weighed, but it was excluded in any statistical calculations as wellas in the calculation of baseline feed intake.

Summary of Findings

As expected, all influenza infected mice developed clinical signs ofdisease starting 2 days post infection. Clinical signs included: 1)ruffled fur, 2) inactivity, 3) hunching, 4) death, 5) weight loss, and6) loss of appetite.

Clinical signs 1-4 were scored by two independent observers using ascale of 0-5. The cumulative scores were plotted and statisticallyanalyzed. Statistical analyses were performed using SAS v.9.1.3. Ananalysis of variance (ANOVA) of a repeated measures design wascalculated in a split-plot arrangement of treatments, with the treatmentgroup on the main plot, and the subplot including day and treatment byday interactions. Interaction of pairwise comparisons of least squaremeans was performed with a pairwise t-test with pre-planned comparisons.This statistical methodology was used for all comparisons in the study.

A. Statistically significant differences were reported in the treatedversus untreated scores in the time interval spanning days 6-8 and postinfection for clinical signs 1-4 (FIG. 3). Specifically, it was notedthat mice individual mice fed with N5G appeared substantially healthierthan control mice as evidenced primarily by ruffled fur, lethargy andhunching posture changes.

B. Measurements of feed intake revealed a statistically significant lossof appetite as revealed by the drop in % daily feed intake especiallynoted in the time span of days 5-6 post infection (FIG. 4).

C. The most dramatic change in treated versus untreated mice was thenoted % change in mouse weights of untreated versus treated (N5G fed)mice. A highly significant reduction in body weights was observed in thecontrol mouse group in the time span between days 3-9 post infection(FIG. 5).

D. The most important aspect of influenza infections is the resultantlung immunopathology. Typically, viral replication as well asinfiltration of lymphocytes causes substantial damage to lung tissues.In fact, it is believed that infiltration of lymphocytes and theirnegative effects on lungs tissues is the most important parameter ofinfluenza-related morbidity and mortality in humans. Consequently,particular care was taken to examine the tissues of mice for signs ofhistopathological changes. The following evaluation criteria wereutilized to score these changes: 1) Lung consolidation scores (LCS); 2)Histopathological scores (HS). LCS were rated as follows: grade 1: noplum coloration; grade 2: minimum plum coloration; grade 3: medium plumcoloration; grade 4: maximum plum coloration. HS were rated as follows:Grade 0: no overt histological change; grade 1+: onlybronchial/bronchiolar and/or peri-bronchial/bronchiolar/vascularchanges; grade 2+: 1-10% pulmonary parenchyma affected; grade 3+: 11-30%pulmonary parenchyma affected; grade 4:+: 31-50% pulmonary parenchymaaffected; grade 5+: greater than 50% pulmonary parenchyma affected.

a. Lung consolidation scores revealed a “trend” toward a healthierstatus of treated versus untreated mice. However, there results were notstatistically significant (FIG. 6).

b. Histopathological scores revealed a statistically significantdifference in treated versus untreated mice at day 6 post infection(FIG. 7).

E. Viral titers were obtained from lung homogenates starting at day 2post infection (days 2, 4, 6, 8 and 21). No significant differences wereobserved in treated versus untreated mice, while viral titers obtainedat 2 days post infection were less than 100 plaque forming units (PFU)per 250iL of homogenate (the lungs were homogenized to produce a 1mL-total homogenate). No statistical analysis could be applied to viraltiters since only one mouse per group per time point was tested.

Overall Conclusion

The obtained results collectively indicate a significant effect of N5Gon ameliorating clinical symptoms and adverse pathological implicationof influenza infection in the mouse experimental model.

All publications and patents mentioned in the above specification areherein incorporated by reference. Various modifications and variationsof the described method and system of the invention will be apparent tothose skilled in the art without departing from the scope and spirit ofthe invention. Although the invention has been described in connectionwith specific preferred embodiments, it should be understood that theinvention as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the invention which are obvious to those skilled in therelevant fields are intended to be within the scope of the followingclaims.

We claim:
 1. A method for alleviating the symptoms of a cold in a humanor animal subject comprising providing a composition comprisingn-glycolylneuraminic acid in a concentration of 1 nM to 10 mM, andintransally administering said composition under conditions such thatsymptoms of a cold are alleviated.
 2. The method of claim 1, whereinsaid composition is selected from the group consisting of gels,solutions, sprays, powders, creams, foams and lotions.